Riboflavin-biosynthesis in fungi

ABSTRACT

The present invention relates to the genes for riboflavin biosynthesis in the fungus Ashbya gossypii and to genetic engineering processes for preparing riboflavin using these genes and gene products.

This application is national stage application of PCT/EP95/00958, filed on Mar. 15, 1995.

The present invention relates to the genes for riboflavin biosynthesis in fungi, to the proteins encoded thereby and to genetic engineering processes for preparing riboflavin using these genes and gene products.

The preparation of riboflavin by fermentation of fungi such as Eremothecium ashbyii or Ashbya gossypii has been disclosed (The Merck Index, Windholz et al., eds. Merck & Co., page 1183 (1983)).

EP 405370 describes riboflavin-overproducing bacterial strains which have been obtained by transformation of the riboflavin bio- synthesis genes from Bacillus subtilis.

Since the genetics of riboflavin biosynthesis in bacteria and eu- karyotes differ, the abovementioned genes from Bacillus subtilis are unsuitable for a recombinant process for preparing riboflavin using eukaryotic producer organisms such as Ashbya gossypii.

The cloning of the riboflavin biosynthesis genes of the yeast Saccharomyces cerevisiae was described in a patent application filed at the German Patent Office on Nov. 19, 1992.

However, it was not possible to clone the Ashbya gossypii ribo- flavin biosynthesis genes using the S. cerevisiae rib genes by conventional hybridization methods; evidently the homology of the rib genes from S. cerevisiae and A. gossypii was not great enough for hybridization.

It is an object of the present invention to isolate the ribo- flavin biosynthesis genes from a eukaryote in order in this way to provide a recombinant process for preparing riboflavin in a eukaryotic producer organism.

We have found that this object is achieved by isolation of six genes (rib genes) found in the ascomycete Ashbya gossypii which code for enzymes of riboflavin biosynthesis starting from GTP.

The invention relates to the following DNA sequences:

DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 2, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 2, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.

DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 4, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 4, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.

DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 6, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 6, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.

DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 8, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 8, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.

DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 10, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 10, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.

DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 12, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 12, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.

The genes and their gene products (polypeptides) are shown in the sequence listing with their primary structure and are assigned as follows:

SEQ ID NO: 1: rib 1 gene

SEQ ID NO: 2: rib 1 gene product (GTP cyclohydrolase II)

SEQ ID NO: 3: rib 2 gene

SEQ ID NO: 4: rib 2 gene product (DRAP deaminase)

SEQ ID NO: 5: rib 3 gene

SEQ ID NO: 6: rib 3 gene product (DBP synthase)

SEQ ID NO: 7: rib 4 gene

SEQ ID NO: 8: rib 4 gene product (DMRL synthase)

SEQ ID NO: 9: rib 5 gene

SEQ ID NO: 10: rib 5 gene product (riboflavin synthase)

SEQ ID NO: 11: rib 7 gene

SEQ ID NO: 12: rib 7 gene product (HTP reductase)

Guanosine triphosphate (GTP) is converted by GTP cyclohydrolase II (rib 1 gene product) into 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5-phosphate. This compound is subsequently reduced by the rib 7 gene product to 2,5-diaminoribitylamino-2,4(1H,3H)-pyrimidine 5-phosphate and then deaminated by the rib 2 gene product to 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione. Subsequently, in a reaction catalyzed by the rib 4 gene product, the C4 compound DBP is added on to result in 6,7-dimethyl-8-ribitvllumazine (DMRL), from which riboflavin is produced in the reaction catalyzed by the rib 5 gene product. The C4 compound DBP (L-3,4-dihydroxy-2-butanone 4-phosphate) is formed from D-ribulose 5-phosphate in a reaction catalyzed by the rib 3 gene product.

The DNA sequences described in SEQ ID NO: 1, 3, 5, 7, 9, 11 code for the polypeptides described in SEQ ID NO: 2, 4, 6, 8, 10, 12.

DNA sequences, apart from those specified in the sequence list- ing, which are also suitable are those which, as a consequence of the degeneracy of the genetic code, have a different DNA sequence but code for the same polypeptide.

The invention also relates to those DNA sequences which code for a gene product (polypeptide) with a primary structure other than that detailed in the sequence listing as long as the gene product still has essentially the same biological properties as the gene product specified in the sequence listing. Biological properties mean, in particular, the enzymatic activities bringing about the biosynthesis of riboflavin.

Such modified gene products with essentially the same biological properties can be obtained by deletion or addition of one or more amino acids or peptides or by replacing amino acids by other ami- no acids, or can be isolated from organisms other than Ashbya gossypii.

The DNA sequences which code for the modified gene products are, as a rule, homologous to the extent of 80 percent or more with the DNA sequences shown in the sequence listing. Such DNA sequences can be isolated starting from the DNA sequences described in SEQ ID NO: 1, 3, 5, 7, 9, 11, for example with conventional hybridization methods or the PCR technique from eukaryotes other than Ashbya gossypii. These DNA sequences hybridize under standard conditions with the DNA sequences described in SEQ ID NO: 1, 3, 5, 7, 9, 11.

Standard conditions mean, for example, temperatures from 42 to 58° C. in an aqueous buffer solution with a concentration of from 0.1 to 1 × SSC (1 × SSC: 0.15 M NaCl, 15 mM sodium citrate pH 7.2). The experimental conditions for DNA hybridizations are described in textbooks of genetic engineering, for example in Sambrook et al., "Molecular Cloning", Cold Spring Harbor Laboratory, 1989.

The invention also relates to regulating sequences, in particular promoter sequences, which are located upstream in the 5' direction of the DNA sequences coding for the appropriate polypeptide. The regulating sequences are specified in the sequence listing and explained in detail below.

Regulating sequence for rib 1 gene: SEQ ID NO: 1 nucleotide 1-242

Regulating sequence for rib 2 gene: SEQ ID NO: 3 nucleotide 1-450

Regulating sequence for rib 3 gene: SEQ ID NO: 5 nucleotide 1-314

Regulating sequence for rib 4 gene: SEQ ID NO: 7 nucleotide 1-270

Regulating sequence for rib 5 gene: SEQ ID NO: 9 nucleotide 1-524

Regulating sequence for rib 7 gene: SEQ ID NO: 11 nucleotide 1-352

The regulating sequences can also be truncated in the 5' and/or 3' direction with negligible diminution in their functioning.

Essential for the regulating action are, as a rule, fragments of 30-100, preferably 40-70, nucleotides from the abovementioned sequence regions.

These regulating sequences can also be optimized in their functioning, by comparison with the natural sequences, by directed mutagenesis.

The regulating sequences according to the invention are suitable for overexpression of genes in Ashbya, in particular of genes responsible for riboflavin biosynthesis.

The invention also relates to expression vectors which contain one or more of the DNA sequences according to the invention. Such expression vectors are obtained by providing the DNA sequences according to the invention with suitable functional regulating signals. Such regulating signals are DNA sequences which are responsible for the expression, for example promoters, operators, enhancers, ribosome binding sites, and which are recognized and obeyed by the host organism.

It is also possible where appropriate for the expression vector to comprise other regulating signals which, for example, control replication or recombination of the recombinant DNA in the host organism.

The invention likewise relates to the host organisms transformed with the DNA sequences or expression vectors according to the invention. Eukaryotic organisms are preferably used as host organisms, particularly preferably those of the genus Saccharomyces, Candida, Pichia, Eremothecium or Ashbya. Particularly preferred species are Saccharomyces cerevisiae, Candida flaveri, Candida famata, Eremothecium ashbyii and Ashbya gossypii.

The invention also includes a recombinant process for preparing riboflavin in which the transformed host organisms according to the invention are cultured in a conventional way by fermentation, and the riboflavin produced during the fermentation is isolated from the fermentation medium and, where appropriate, purified.

The rib genes and gene products can be isolated and characterized as described in the example and in the sequence listing.

EXAMPLE 1

Isolation of the Ashbya gossypii riboflavin biosynthesis genes (rib genes)

a. Construction of an Ashbya gossypii cDNA bank

RNA was completely extracted from the mycelium of the riboflavin-overproducing strain Ashbya gossypii ATCC 10195 in the late logarithmic phase of growth after cultivation on YEPD medium (Sherman et al., "Methods in yeast genetics", Cold Spring Harbor, N.Y., 1989).

Poly(A)⁺ RNA was purified by adsorption on and elution from oligo(dT)-cellulose twice (Aviv and Leder, Proc. Natl. Acad. Sci. USA 69, 1972, 1408-1412). The cDNA was isolated by the general method of Gubler and Hoffmann (Gene 25, 1983, 263), and synthetic EcoRI adaptors were added on to the ends of the blunt-ended cDNA molecules. The cDNA fragments after cutting with EcoRI were subsequently phosphorylated using T4 polynucleotide kinase and cloned into the dephosphorylated vector pYEura3 which had been cut with EcoRI (FIG. 1). pYEura3 (Clonetech Laboratories, Inc., Calif.) is a yeast expression vector which contains the galactose-inducible GALL and GALIO promoters and URA, CEN4 and ARS1. These yeast elements permit the transformation and expres- sion of cloned DNA fragments in yeast cells.

Aliquots from the ligation reaction were used to transform highly competent (Hanahan, DNA Cloning, ed. D.M. Glover; IRL Press, oxford 1985, 109) E. coli XL1-Blue (Bullock et al., Biotechniques 5 (1987) 376-378), and transformants were selected on the basis of their ampicillin resistance.

About 3×10⁵ ampicillin-resistant cells were combined and ampli- fied, and plasmid DNA was isolated therefrom (Birnboim and Doly, Nucleic Acids Res. 7, 1979, 1513).

b. Isolation of Ashbya gossypii cDNA clones which code for riboflavin-producing enzymes

cDNA clones from Ashbya gossypii which code for riboflavin- producing enzymes were isolated by functional complementation of Saccharomyces cerevisiae mutants involved in riboflavin biosynthesis.

The strains AJ88 (Mata leu2 his3 rib1::URA3 ura3-52), AJ115 (Matalpha leu2 inos1 rib2::URA3 ura3-52), AJ71 (Matalpha leu2 inos1 rib3::URA3 ura3-52), AJ106 (Matalpha leu2 inos1 rib4::URA3 ura3-52), AJ66 (Mata canR inos1 rib5::URA3 ura3-52) and AJ121 (Matalpha leu2 inos1 rib7::URA3 ura3-52) are mutated strains produced by destruction of one of the six genes (rib1 to rib5 and rib7) involved in riboflavin biosynthesis in Saccharomyces cerevisiae.

These strains were each transformed with 25 μg of cDNA from the Ashbya gossypii CDNA bank and plated on solid galactose-containing medium without riboflavin. After growth for approximately one week, rib⁺ transformants were isolated from the culture dishes.

In each case one transformant from each transformed mutant (Rib1+, Rib2+, Rib3+, Rib4+, Rib5+ and Rib7+) was analyzed and it was found in all cases that the Rib⁺ phenotype was expressed only in galactose medium but not in glucose medium.

These results demonstrate that the Rib+ phenotype was expressed under the control of the galactose-inducible GAL10 promoter located on the plasmid.

Plasmid DNA was isolated from the Rib1+, Rib2+, Rib3+, Rib4+, Rib5+ and Rib7+ transformants by transformation of E. coli and was called pJR715, pJR669, pJR788, pJR733, pJR681 and pJR827.

Partial sequencing of the cDNA inserts present in these plasmids confirmed that they code for proteins which are analogous to proteins of the rib gene products from Saccharomyces.

c. Isolation of Ashbya gossypii genomic clones which code for ri- boflavin-producing enzymes

In order to isolate the genomic copies of the riboflavin-producing genes of Ashbya gossypii, a genomic bank of Ashbya gossypii ATCC 10195 was constructed in the cosmid supercos1 (Stratagene Cloning Systems, Calif.) and screened with ³² P-labeled probes which were derived from the cDNA copies of the rib1, rib2, rib3, rib4, rib5 and rib7 genes of Ashbya gossypii.

Cosmid clones with rib1, rib2, rib3, rib4, rib5 and rib7 DNA were isolated by colony hybridization (Grunstein and Hogness, Proc. Natl. Acad. Sci. USA 72, 1975, 3961-3965). Further Southern analyses of enzymatically cleaved cosmid DNA using the same rib-specific cDNA probes made it possible to identify defined restriction fragments which contained the rib1, rib2, rib3, rib4, rib5 and rib7 genes of Ashbya gossypii.

A BamHI-ClaI DNA fragment which was 3.1 kb in length and contained the complete rib1 gene of Ashbya gossypii coding for GTP cyclohydrolase II was found. This fragment was isolated from an agarose gel and cloned into the pBluescript KS (+) phagemid (Stratagene Cloning Systems) cut with BamHI and ClaI, and in this way provided the plasmid pJR765 (FIG. 2).

A DNA sequence (SEQ ID NO: 1) which was 1,329 bp in length and contained the rib1 open reading frame of 906 bp, 242 bp of the 5'-noncoding region and 181 bp of the 3'-noncoding region was ob- tained.

The complete Ashbya gossypii rib2 gene which codes for DRAP deaminase was found on an EcoRI-PstI fragment which was 3.0 kb in length and which, cloned into pBluescript KS (+), yielded the plasmid pJR758 (FIG. 3).

A region 2,627 bp in length from the EcoRI-PstI insert with the 1,830 bp open reading frame of rib2, 450 bp of the 5'-untranslated region and 347 bp of the 3'-untranslated region was sequenced (SEQ ID NO: 3).

The complete Ashbya gossypii rib 3 gene which codes for DBP synthase was found on a PstI-HindIII fragment which was 1.5 kb in length and, cloned into pBluescript KS (+), yielded the plasmid pJR790 (FIG. 4).

A region 1,082 bp in length from the PstI-HindIII insert with the 639 bp open reading frame of rib 3, 314 bp of the 5'-untranslated region and 129 bp of the 3'-untranslated region was sequenced (SEQ ID NO: 5).

The Ashbya gossypii rib4 gene which codes for DMRL synthase was found on a PstI-PstI fragment which was 3.2 kb in length and, cloned into pBluescript KS (+), yielded the plasmid pJR762 (FIG. 5).

A region 996 bp in length from the PstI-PstI insert with the 519 bp open reading frame of rib4, 270 bp of the 5'-untranslated region and 207 bp of the 3'-untranslated region was sequenced (SEQ ID NO: 7).

The complete Ashbya gossypii rib5 gene which codes for riboflavin synthase was found on a PstI-PstI fragment which was 2.5 kb in length and, cloned into pBluescript KS (+), yielded the plasmid pJR739 (FIG. 6).

A region 1,511 bp in length from the PstI-PstI insert with the 708 bp open reading frame of rib5, 524 bp of the 5'-untranslated region and 279 bp of the 3'-untranslated region was sequenced (SEQ ID NO: 9).

Finally, the Ashbya gossypii rib7 gene which codes for HTP reductase was found on an EcoRI-EcoRI fragment which was 4.1 kb in length and, cloned into pBluescript KS (+), yielded the plasmid pJR845 (FIG. 7).

A region 1,596 bp in length from the EcoRI-EcoRI insert with the 741 bp open reading frame of rib7, 352 bp of the 5'-untranslated region and 503 bp of the 3'-untranslated region was sequenced (SEQ ID NO: 11).

EXAMPLE 2

mRNA analysis of the Ashbya gossypii rib genes

Northern analyses were carried out to identify the rib-specific transcripts. The total RNA was isolated from the Ashbya gossypii strain ATCC 10195 as described in Example 1. The RNA samples from the strain (5 μg) were fractionated by electrophoresis on 0.8% agarose formaldehyde gels together with RNA size markers and blotted in vacuo onto nylon membranes (Thomas, Proc. Natl. Acad. Sci. USA, 77, 1980, 5201-5205).

The nylon membranes were separately hybridized with ³² P-labeled rib-specific DNA probes at 42° C. in 5 × SSC and in the presence of 50% formamide. The Ashbya gossypii rib1 gene is expressed as unique message of about 1,150 nucleotides, which was detected in both strains by an SmaI-SacI probe 0.7 kbp in length from the plasmid pJR765 (FIG. 8).

In a similar way, unique 1,900 nucleotide-long rib2, 900 nucleotide-long rib3, 800 nucleotide-long rib4, 1,050 nucleotidelong rib5 and 1,000 nucleotide-long rib7 transcripts were detected in the blots using an SmaI-SmaI fragment 0.5 kbp in length from pJR758, a HindIII-KpnI fragment 0.6 kbp in length from pJR790, an ScaI-HindIII fragment 0.5 kbp in length from pJR739 and a PstI-PstI fragment 0.3 kbp in length from pJR845 as specific probe.

EXAMPLE 3

Expression of the Ashbya gossypii rib genes in Saccharomyces cerevisiae

It is possible as described in Example 1 to grow Saccharomyces cerevisiae mutants which have been well investigated and are defective in one stage of riboflavin biosynthesis on culture media without riboflavin if they harbor a plasmid which codes for the complementing Ashbya enzymes. In order to test the function of the Ashbya gossypii rib gene products, flavin-producing enzyme activities were measured in cell-free extracts from S. cerevisiae mutants which harbored one of the expression plasmids pJR715, pJR669, pJR788, pJR733, pJR681 and pJR827.

These plasmids which are derived from pYEura3 and are described in Example 1 contain Ashbya gossypii rib-specific cDNA fragments under the control of the galactose-inducible GAL10 promoter.

Cell-free protein extracts from S. cerevisiae were obtained from cultures which had grown in liquid medium to an optical density of about 2 OD.

The cells were harvested, washed with cold 20 mM tris HCl, pH 7.5, and resuspended in the same buffer, which was supplemented with 1 mM phenylethylsulfonyl fluoride.

Cell lysates were prepared by vortexing in the presence of glass beads and centrifuging at 3,000 g and 4° C. for 20 min.

GTP cyclohydrolase II, DRAP deaminase, DBP synthase, DMRL synthase, riboflavin synthase and HTP reductase enzyme activities were determined as described in the literature (Shavlovsky et al., Arch. Microbiol. 124, 1980, 255-259; Richter et al., J. Bacteriol. 175, 1993, 4045-4051; Klein and Bacher, Z. Naturforsch. 35b, 1980, 482-484; Richter et al., J. Bacteriol. 174, 1992, 4050-4056; Nielsen et al., J. Biol. Chem. 261, 1986, 3661; Plaut and Harvey, Methods Enzymol. 18B, 1971, 515-538; Hollander and Brown, Biochem. Biophys. Res. Commun. 89, 1979, 759-763; Shavlovski et al., Biochim. Biophys. Acta, 428, 1976, 611-618).

Protein was quantified by the Peterson method (Anal. Biochem. 83, 1977, 346-356). As is evident from Tab. 1, the plasmid pJR715 brings about the expression of GTP cyclohydrolase II activity in the S. cerevisiae mutant AJ88. Furthermore, this activity is present only in cells which have grown on galactose medium, which indicates that the rib1 cDNA expression of Ashbya gossypii takes place under the control of the galactose-inducible GAL10 promoter.

These results therefore demonstrate that rib1 codes for GTP cyclohydrolase II in Ashbya gossypii. It was shown in a similar way that rib2 codes for DRAP deaminase, rib3 codes for DBP synthase, rib4 codes for DMRL synthase, rib5 codes for riboflavin synthase and rib7 codes for HTP reductase in this fungus.

                  TABLE 1                                                          ______________________________________                                         GTP cyclohydrolase II activity of the S. cerevisiae ribl mutant                AJ88 and its transformants.                                                                           GTP cyclohydrolase II                                                          U/mg protein **)                                        Strain     Plasmid     Glucose    Galactose                                    ______________________________________                                         X 2180-1A* --          0.48       0.34                                         AJ 88      --          n.d.       n.d.                                         AJ 88      pIR715      n.d.       21.60                                        ______________________________________                                          n.d.: not detected                                                             *) Wildtype                                                                    **) Units of GTP cyclohydrolase II activities 1U catalyzes the formation       of 1 nmol of HTP per hour                                                

                  TABLE 2                                                          ______________________________________                                         DRAP deaminase activity of the S. cerevisiae rib2 mutant AJ115                 and its transformants.                                                                                DRAP deaminase                                                                 U/mg protein *)                                         Strain     Plasmid     Glucose   Galactose                                     ______________________________________                                         X 2180-1A  --          0.45      0.38                                          AJ 115     --          n.d.      n.d.                                          AJ 115     pIR669      n.d.      53.22                                         ______________________________________                                          n.d.: not detected                                                             *) 1U catalyzes the formation of 1 nmol of ARAP per hour                 

                  TABLE 3                                                          ______________________________________                                         DBP synthase activity of the S. cerevisiae rib3 mutant AJ71                    and its transformants.                                                                                DBP synthase                                                                   U/mg protein *)                                         Strain     Plasmid     Glucose   Galactose                                     ______________________________________                                         X 2180-1A  --          0.80      0.75                                          AJ 71      --          n.d.      n.d.                                          AJ 71      pIR788      n.d.      25.19                                         ______________________________________                                          n.d.: not detected                                                             *) 1U catalyzes the formation of 1 nmol of DBP per hour                  

                  TABLE 4                                                          ______________________________________                                         DMRL synthase activity of the S. cerevisiae rib4 mutant AJ106                  and its transformants.                                                                                DMRL synthase                                                                  U/mg protein *)                                         Strain     Plasmid     Glucose   Galactose                                     ______________________________________                                         X 2180-1A  --          2.04      1.73                                          AJ 106     --          n.d.      n.d.                                          AJ 106     pIR733      n.d.      86.54                                         ______________________________________                                          n.d.: not detected                                                             *) 1U catalyzes the formation of 1 nmol of DMRL per hour                 

                  TABLE 5                                                          ______________________________________                                         Riboflavin synthase activity of the S. cerevisiae rib5 mutant                  AJ66 and its transformants.                                                                           Riboflavin synthase                                                            U/mg protein *)                                         Strain     Plasmid     Glucose   Galactose                                     ______________________________________                                         X 2180-1A  --          4.41      3.80                                          AJ 66      --          n.d.      n.d.                                          AJ 66      pIR681      n.d.      164.20                                        ______________________________________                                          n.d.: not detected                                                             *) 1U catalyzes the formation of 1 nmol of riboflavin per hour           

                  TABLE 6                                                          ______________________________________                                         HTP reductase activity of the S. cerevisiae rib7 mutant AJ121                  and its transformants.                                                                                HTP reductase                                                                  U/mg protein *)                                         Strain     Plasmid     Glucose   Galactose                                     ______________________________________                                         X 2180-1A  --          1.86      2.54                                          AJ 121     --          n.d.      n.d.                                          AJ 121     pIR827      n.d.      46.21                                         ______________________________________                                          n.d.: not detected                                                             *) 1U catalyzes the formation of 1 nmol of DRAP per hour                 

    __________________________________________________________________________     SEQUENCE LISTING                                                               (1) GENERAL INFORMATION:                                                       (iii) NUMBER OF SEQUENCES: 12                                                  (2) INFORMATION FOR SEQ ID NO: 1:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 1329 base pairs                                                    (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: cDNA to mRNA                                               (iii) HYPOTHETICAL: NO                                                         (iii) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Ashbya gossypii                                                  (ix) FEATURE:                                                                  (A) NAME/KEY: 5'UTR                                                            (B) LOCATION: 1..242                                                           (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                              (B) LOCATION: 243..1148                                                        (ix) FEATURE:                                                                  (A) NAME/KEY: 3'UTR                                                            (B) LOCATION: 1149..1329                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:                                       TTTCTGTCCGCATACTTCATATGCTCATCGCACATTGATAATGTACATTCGAAAAATTTC60                 AAGATTAGCCTCCGTGAACAGCGATTTACCTTAGGCAAAAGTAACAAAAGGCTTTTCCGT120                AGGTGCTTTGTCATTCAACAATCCACGTCGGAATTGGCGACTATATAGTGTAGGGCCCAT180                AAAGCAGTAGTCGGTGTTGATAGCTGTGTCAGACCAACTCTTTGTTAATTACTGAAGCTG240                ATATGACTGAATACACAGTGCCAGAAGTGAGGTGTGTCGCACGCGCG287                             MetThrGluTyrThrValProGluValArgCysValAlaArgAla                                  151015                                                                         CGCATACCGACGGTACAGGGCACCGATGTCTTCCTCCATCTATACCAC335                            ArgIleProThrValGlnGlyThrAspValPheLeuHisLeuTyrHis                               202530                                                                         AACTCGATCGACAGCAAGGAACACCTAGCGATTGTCTTCGGCGAGAAC383                            AsnSerIleAspSerLysGluHisLeuAlaIleValPheGlyGluAsn                               354045                                                                         ATACGCTCGCGGAGTCTGTTCCGGTACCGGAAAGACGACACGCAGCAG431                            IleArgSerArgSerLeuPheArgTyrArgLysAspAspThrGlnGln                               505560                                                                         GCGCGGATGGTGCGGGGCGCCTACGTGGGCCAGCTGTACCCCGGGCGG479                            AlaArgMetValArgGlyAlaTyrValGlyGlnLeuTyrProGlyArg                               657075                                                                         ACCGAGGCAGACGCGGATCGGCGTCAGGGCCTGGAGCTGCGGTTTGAT527                            ThrGluAlaAspAlaAspArgArgGlnGlyLeuGluLeuArgPheAsp                               80859095                                                                       GAGACAGGGCAGCTGGTGGTGGAGCGGGCGACGACGTGGACCAGGGAG575                            GluThrGlyGlnLeuValValGluArgAlaThrThrTrpThrArgGlu                               100105110                                                                      CCGACACTGGTGCGGCTGCACTCGGAGTGTTACACGGGCGAGACGGCG623                            ProThrLeuValArgLeuHisSerGluCysTyrThrGlyGluThrAla                               115120125                                                                      TGGAGCGCGCGGTGCGACTGCGGGGAGCAGTTCGACCAGGCGGGTAAG671                            TrpSerAlaArgCysAspCysGlyGluGlnPheAspGlnAlaGlyLys                               130135140                                                                      CTGATGGCTGCGGCGACAGAGGGCGAGGTGGTTGGCGGTGCGGGGCAC719                            LeuMetAlaAlaAlaThrGluGlyGluValValGlyGlyAlaGlyHis                               145150155                                                                      GGCGTGATCGTGTACCTGCGGCAGGAGGGCCGCGGCATCGGGCTAGGC767                            GlyValIleValTyrLeuArgGlnGluGlyArgGlyIleGlyLeuGly                               160165170175                                                                   GAGAAGCTGAAGGCGTACAACCTGCAGGACCTGGGCGCGGACACGGTG815                            GluLysLeuLysAlaTyrAsnLeuGlnAspLeuGlyAlaAspThrVal                               180185190                                                                      CAGGCGAACGAGCTGCTCAACCACCCTGCGGACGCGCGCGACTTCTCG863                            GlnAlaAsnGluLeuLeuAsnHisProAlaAspAlaArgAspPheSer                               195200205                                                                      TTGGGGCGCGCAATCCTACTGGACCTCGGTATCGAGGACATCCGGTTG911                            LeuGlyArgAlaIleLeuLeuAspLeuGlyIleGluAspIleArgLeu                               210215220                                                                      CTCACGAATAACCCCGACAAGGTGCAGCAGGTGCACTGTCCGCCGGCG959                            LeuThrAsnAsnProAspLysValGlnGlnValHisCysProProAla                               225230235                                                                      CTACGCTGCATCGAGCGGGTGCCCATGGTGCCGCTTTCATGGACTCAG1007                           LeuArgCysIleGluArgValProMetValProLeuSerTrpThrGln                               240245250255                                                                   CCCACACAGGGCGTGCGCTCGCGCGAGCTGGACGGCTACCTGCGCGCC1055                           ProThrGlnGlyValArgSerArgGluLeuAspGlyTyrLeuArgAla                               260265270                                                                      AAGGTCGAGCGCATGGGGCACATGCTGCAGCGGCCGCTGGTGCTGCAC1103                           LysValGluArgMetGlyHisMetLeuGlnArgProLeuValLeuHis                               275280285                                                                      ACGTCTGCGGCGGCCGAGCTCCCCCGCGCCAACACACACATATAATCTTTGC1155                       ThrSerAlaAlaAlaGluLeuProArgAlaAsnThrHisIle                                     290295300                                                                      TATATTAAAACTCTATAAACGTATGCCACACGGCGCCCGCGGGCTGCCACACGCTGCTCA1215               CGGGCTGCCGAACAGTTCTAACAAGTAATCGCGCGCCTCGCCAGTGATCGTGGCGAGCAC1275               CTTGTCGTCCATCATCACATATCCTCGGCTACAGTCGTCGTTGAAGAGCGTGCA1329                     (2) INFORMATION FOR SEQ ID NO: 2:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 301 amino acids                                                    (B) TYPE: amino acid                                                           (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: protein                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:                                       MetThrGluTyrThrValProGluValArgCysValAlaArgAlaArg                               151015                                                                         IleProThrValGlnGlyThrAspValPheLeuHisLeuTyrHisAsn                               202530                                                                         SerIleAspSerLysGluHisLeuAlaIleValPheGlyGluAsnIle                               354045                                                                         ArgSerArgSerLeuPheArgTyrArgLysAspAspThrGlnGlnAla                               505560                                                                         ArgMetValArgGlyAlaTyrValGlyGlnLeuTyrProGlyArgThr                               65707580                                                                       GluAlaAspAlaAspArgArgGlnGlyLeuGluLeuArgPheAspGlu                               859095                                                                         ThrGlyGlnLeuValValGluArgAlaThrThrTrpThrArgGluPro                               100105110                                                                      ThrLeuValArgLeuHisSerGluCysTyrThrGlyGluThrAlaTrp                               115120125                                                                      SerAlaArgCysAspCysGlyGluGlnPheAspGlnAlaGlyLysLeu                               130135140                                                                      MetAlaAlaAlaThrGluGlyGluValValGlyGlyAlaGlyHisGly                               145150155160                                                                   ValIleValTyrLeuArgGlnGluGlyArgGlyIleGlyLeuGlyGlu                               165170175                                                                      LysLeuLysAlaTyrAsnLeuGlnAspLeuGlyAlaAspThrValGln                               180185190                                                                      AlaAsnGluLeuLeuAsnHisProAlaAspAlaArgAspPheSerLeu                               195200205                                                                      GlyArgAlaIleLeuLeuAspLeuGlyIleGluAspIleArgLeuLeu                               210215220                                                                      ThrAsnAsnProAspLysValGlnGlnValHisCysProProAlaLeu                               225230235240                                                                   ArgCysIleGluArgValProMetValProLeuSerTrpThrGlnPro                               245250255                                                                      ThrGlnGlyValArgSerArgGluLeuAspGlyTyrLeuArgAlaLys                               260265270                                                                      ValGluArgMetGlyHisMetLeuGlnArgProLeuValLeuHisThr                               275280285                                                                      SerAlaAlaAlaGluLeuProArgAlaAsnThrHisIle                                        290295300                                                                      (2) INFORMATION FOR SEQ ID NO: 3:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 2627 base pairs                                                    (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: cDNA to mRNA                                               (iii) HYPOTHETICAL: NO                                                         (iii) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Ashbya gossypii                                                  (ix) FEATURE:                                                                  (A) NAME/KEY: 5'UTR                                                            (B) LOCATION: 1..450                                                           (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                              (B) LOCATION: 451..2280                                                        (ix) FEATURE:                                                                  (A) NAME/KEY: 3'UTR                                                            (B) LOCATION: 2281..2627                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:                                       CTGCAGGACAATTTAAATTACGATTACACGCGGCAGCCTTCTTGGTGCGACAGGATTTTG60                 TACAAGAATGACCCCAAGCGGGTAAGAGTTCATAGGTATGCCTCGATTGATAGACGTTCC120                ATTTTGAATTATACTGATCACGAACCCGTAACGCTCGATGTCAGCGTTTCATGCCATACA180                CAATTTGTCCCAATGGCTATGCAGAATATTTCCCCACAGAGCACCATGGAAATGTATGTG240                GGAGACGTCACAGATATACTACTGATGTTGTTCTCCAGAGTATACTACGCCCCTACCATA300                TTCGATCTTGTGGTATTGACGATATTCCTCTGTTTGGTTTTACTGGCACTATTCCGTTTG360                ACGGTATAGCGCTATTCGTTCATAGTGACACATGCGGCACTAGCTATTCAGCGAATCCTT420                TATAAACTGCTACTTAACGTTCGTAACACCATGCTCAAAGGCGTTCCTGGCCTT474                      MetLeuLysGlyValProGlyLeu                                                       15                                                                             CTTTTTAAGGAGACGCAACGTCATCTGAAACCCAGGCTGGTTAGGATT522                            LeuPheLysGluThrGlnArgHisLeuLysProArgLeuValArgIle                               101520                                                                         ATGGAAAACACATCGCAGGATGAGAGTCGCAAAAGACAGGTCGCTTCG570                            MetGluAsnThrSerGlnAspGluSerArgLysArgGlnValAlaSer                               25303540                                                                       AACTTGAGCAGCGATGCCGATGAGGGCTCGCCGGCAGTTACGAGGCCG618                            AsnLeuSerSerAspAlaAspGluGlySerProAlaValThrArgPro                               455055                                                                         GTTAAAATCACCAAACGCCTCAGGAAGAAGAACCTCGGGACAGGCGAG666                            ValLysIleThrLysArgLeuArgLysLysAsnLeuGlyThrGlyGlu                               606570                                                                         CTACGGGACAAAGCAGGATTCAAGTTGAAGGTGCAAGACGTGAGCAAA714                            LeuArgAspLysAlaGlyPheLysLeuLysValGlnAspValSerLys                               758085                                                                         AACCGTCACAGACAGGTCGATCCGGAATACGAAGTCGTGGTAGATGGC762                            AsnArgHisArgGlnValAspProGluTyrGluValValValAspGly                               9095100                                                                        CCGATGCGCAAGATCAAACCGTATTTCTTCACATACAAGACTTTCTGC810                            ProMetArgLysIleLysProTyrPhePheThrTyrLysThrPheCys                               105110115120                                                                   AAGGAGCGCTGGAGAGATCGGAAGTTGCTTGATGTGTTTGTGGATGAA858                            LysGluArgTrpArgAspArgLysLeuLeuAspValPheValAspGlu                               125130135                                                                      TTTCGGGACCGCGATAGGCCTTACTACGAGAAAGTCATCGGTTCGGGT906                            PheArgAspArgAspArgProTyrTyrGluLysValIleGlySerGly                               140145150                                                                      GGTGTGCTCCTGAACGGTAAGTCATCGACGTTAGATAGCGTATTGCGT954                            GlyValLeuLeuAsnGlyLysSerSerThrLeuAspSerValLeuArg                               155160165                                                                      AATGGAGACCTCATTTCGCACGAGCTGCACCGTCATGAGCCACCGGTC1002                           AsnGlyAspLeuIleSerHisGluLeuHisArgHisGluProProVal                               170175180                                                                      TCCTCTAGGCCGATTAGGACGGTGTACGAAGATGATGACATCCTGGTG1050                           SerSerArgProIleArgThrValTyrGluAspAspAspIleLeuVal                               185190195200                                                                   ATTGACAAGCCCAGCGGGATTCCAGCCCATCCCACCGGGCGTTACCGC1098                           IleAspLysProSerGlyIleProAlaHisProThrGlyArgTyrArg                               205210215                                                                      TTCAACTCCATTACGAAAATACTTGAAAAACAGCTTGGATACACTGTT1146                           PheAsnSerIleThrLysIleLeuGluLysGlnLeuGlyTyrThrVal                               220225230                                                                      CATCCATGTAACCGACTGGACCGCCTAACCAGTGGCCTAATGTTCTTG1194                           HisProCysAsnArgLeuAspArgLeuThrSerGlyLeuMetPheLeu                               235240245                                                                      GCAAAAACTCCAAAGGGAGCCGATGAGATGGGTGATCAGATGAAGGCG1242                           AlaLysThrProLysGlyAlaAspGluMetGlyAspGlnMetLysAla                               250255260                                                                      CGCGAAGTGAAGAAAGAATATGTTGCCCGGGTTGTTGGGGAATTTCCT1290                           ArgGluValLysLysGluTyrValAlaArgValValGlyGluPhePro                               265270275280                                                                   ATAGGTGAGATAGTTGTGGATATGCCACTGAAGACTATAGAGCCGAAG1338                           IleGlyGluIleValValAspMetProLeuLysThrIleGluProLys                               285290295                                                                      CTTGCCCTAAACATGGTTTGCGACCCGGAAGACGAAGCGGGCAAGGGC1386                           LeuAlaLeuAsnMetValCysAspProGluAspGluAlaGlyLysGly                               300305310                                                                      GCTAAGACGCAGTTCAAAAGAATCAGCTACGATGGACAAACGAGCATA1434                           AlaLysThrGlnPheLysArgIleSerTyrAspGlyGlnThrSerIle                               315320325                                                                      GTCAAGTGCCAACCGTACACGGGCCGGACGCATCAGATCCGTGTTCAC1482                           ValLysCysGlnProTyrThrGlyArgThrHisGlnIleArgValHis                               330335340                                                                      TTGCAATACCTGGGCTTCCCAATTGCCAACGATCCGATTTATTCCAAT1530                           LeuGlnTyrLeuGlyPheProIleAlaAsnAspProIleTyrSerAsn                               345350355360                                                                   CCGCACATATGGGGCCCAAGTCTGGGCAAGGAATGCAAAGCAGACTAC1578                           ProHisIleTrpGlyProSerLeuGlyLysGluCysLysAlaAspTyr                               365370375                                                                      AAGGAGGTCATCCAAAAACTAAACGAAATTGGTAAGACTAAATCTGCG1626                           LysGluValIleGlnLysLeuAsnGluIleGlyLysThrLysSerAla                               380385390                                                                      GAAAGTTGGTACCATTCTGATTCCCAAGGTGAAGTTTTCAAAGGGGAA1674                           GluSerTrpTyrHisSerAspSerGlnGlyGluValPheLysGlyGlu                               395400405                                                                      CAATGCGATGAATGTGGCACCGAACTGTACACTGACCCGGGCCCGAAT1722                           GlnCysAspGluCysGlyThrGluLeuTyrThrAspProGlyProAsn                               410415420                                                                      GATCTTGACTTATGGTTGCATGCATATCGGTATGAATCCACTGAACTG1770                           AspLeuAspLeuTrpLeuHisAlaTyrArgTyrGluSerThrGluLeu                               425430435440                                                                   GATGAGAACGGTGCTAAAAAGCGGAGTTACTCTACTGCGTTTCCTGAG1818                           AspGluAsnGlyAlaLysLysArgSerTyrSerThrAlaPheProGlu                               445450455                                                                      TGGGCTCTTGAGCAGCACGGCGACTTCATGCGGCTTGCCATCGAACAG1866                           TrpAlaLeuGluGlnHisGlyAspPheMetArgLeuAlaIleGluGln                               460465470                                                                      GCTAAGAAATGCCCACCCGCGAAGACATCATTTAGCGTTGGTGCCGTG1914                           AlaLysLysCysProProAlaLysThrSerPheSerValGlyAlaVal                               475480485                                                                      TTAGTTAATGGGACCGAGATTTTGGCCACTGGTTACTCACGGGAGCTG1962                           LeuValAsnGlyThrGluIleLeuAlaThrGlyTyrSerArgGluLeu                               490495500                                                                      GAAGGCAACACGCACGCTGAACAATGTGCACTTCAAAAATATTTTGAA2010                           GluGlyAsnThrHisAlaGluGlnCysAlaLeuGlnLysTyrPheGlu                               505510515520                                                                   CAACATAAAACCGACAAGGTTCCTATTGGTACAGTAATATACACGACT2058                           GlnHisLysThrAspLysValProIleGlyThrValIleTyrThrThr                               525530535                                                                      ATGGAGCCTTGTTCTCTCCGTCTCAGTGGTAATAAACCGTGTGTTGAG2106                           MetGluProCysSerLeuArgLeuSerGlyAsnLysProCysValGlu                               540545550                                                                      CGTATAATCTGCCAGCAGGGTAATATTACTGCTGTTTTTGTTGGCGTA2154                           ArgIleIleCysGlnGlnGlyAsnIleThrAlaValPheValGlyVal                               555560565                                                                      CTTGAGCCAGACAACTTCGTGAAGAACAATACAAGTCGTGCGCTATTG2202                           LeuGluProAspAsnPheValLysAsnAsnThrSerArgAlaLeuLeu                               570575580                                                                      GAACAACATGGTATAGACTATATTCTTGTCCCTGGGTTTCAAGAAGAA2250                           GluGlnHisGlyIleAspTyrIleLeuValProGlyPheGlnGluGlu                               585590595600                                                                   TGTACTGAAGCCGCATTGAAGGGTCATTGATTTTGCTGCGAATTGTA2297                            CysThrGluAlaAlaLeuLysGlyHis                                                    605610                                                                         GATGACTTAAAATATCGAGGCGTATAATTCGTCGCATTTTATATAGTTATCTATGTTTAC2357               ATGACTGTTTAAGCTTGATCTATATTTCTCAAGTGAATTGCCACATATGTTGGTACGGTA2417               ATAAATTAATGAGGGAGTTTTGAAATTCGCAACCAATCTTATATACGTTTGATGATATAA2477               ACGGATTGAGATTCATTAAGCTACCTGATTTTCGCTGAACTGTTTGTTATAGGTTTTTAC2537               AGTAAGATAGTTCCTAAGTTTGTTTATTGTCCCCAGTCGGCCAATTGTTCCGGACTTATT2597               ATTATTACCATTAGTGGTGTTAGTAGTATT2627                                             (2) INFORMATION FOR SEQ ID NO: 4:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 609 amino acids                                                    (B) TYPE: amino acid                                                           (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: protein                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:                                       MetLeuLysGlyValProGlyLeuLeuPheLysGluThrGlnArgHis                               151015                                                                         LeuLysProArgLeuValArgIleMetGluAsnThrSerGlnAspGlu                               202530                                                                         SerArgLysArgGlnValAlaSerAsnLeuSerSerAspAlaAspGlu                               354045                                                                         GlySerProAlaValThrArgProValLysIleThrLysArgLeuArg                               505560                                                                         LysLysAsnLeuGlyThrGlyGluLeuArgAspLysAlaGlyPheLys                               65707580                                                                       LeuLysValGlnAspValSerLysAsnArgHisArgGlnValAspPro                               859095                                                                         GluTyrGluValValValAspGlyProMetArgLysIleLysProTyr                               100105110                                                                      PhePheThrTyrLysThrPheCysLysGluArgTrpArgAspArgLys                               115120125                                                                      LeuLeuAspValPheValAspGluPheArgAspArgAspArgProTyr                               130135140                                                                      TyrGluLysValIleGlySerGlyGlyValLeuLeuAsnGlyLysSer                               145150155160                                                                   SerThrLeuAspSerValLeuArgAsnGlyAspLeuIleSerHisGlu                               165170175                                                                      LeuHisArgHisGluProProValSerSerArgProIleArgThrVal                               180185190                                                                      TyrGluAspAspAspIleLeuValIleAspLysProSerGlyIlePro                               195200205                                                                      AlaHisProThrGlyArgTyrArgPheAsnSerIleThrLysIleLeu                               210215220                                                                      GluLysGlnLeuGlyTyrThrValHisProCysAsnArgLeuAspArg                               225230235240                                                                   LeuThrSerGlyLeuMetPheLeuAlaLysThrProLysGlyAlaAsp                               245250255                                                                      GluMetGlyAspGlnMetLysAlaArgGluValLysLysGluTyrVal                               260265270                                                                      AlaArgValValGlyGluPheProIleGlyGluIleValValAspMet                               275280285                                                                      ProLeuLysThrIleGluProLysLeuAlaLeuAsnMetValCysAsp                               290295300                                                                      ProGluAspGluAlaGlyLysGlyAlaLysThrGlnPheLysArgIle                               305310315320                                                                   SerTyrAspGlyGlnThrSerIleValLysCysGlnProTyrThrGly                               325330335                                                                      ArgThrHisGlnIleArgValHisLeuGlnTyrLeuGlyPheProIle                               340345350                                                                      AlaAsnAspProIleTyrSerAsnProHisIleTrpGlyProSerLeu                               355360365                                                                      GlyLysGluCysLysAlaAspTyrLysGluValIleGlnLysLeuAsn                               370375380                                                                      GluIleGlyLysThrLysSerAlaGluSerTrpTyrHisSerAspSer                               385390395400                                                                   GlnGlyGluValPheLysGlyGluGlnCysAspGluCysGlyThrGlu                               405410415                                                                      LeuTyrThrAspProGlyProAsnAspLeuAspLeuTrpLeuHisAla                               420425430                                                                      TyrArgTyrGluSerThrGluLeuAspGluAsnGlyAlaLysLysArg                               435440445                                                                      SerTyrSerThrAlaPheProGluTrpAlaLeuGluGlnHisGlyAsp                               450455460                                                                      PheMetArgLeuAlaIleGluGlnAlaLysLysCysProProAlaLys                               465470475480                                                                   ThrSerPheSerValGlyAlaValLeuValAsnGlyThrGluIleLeu                               485490495                                                                      AlaThrGlyTyrSerArgGluLeuGluGlyAsnThrHisAlaGluGln                               500505510                                                                      CysAlaLeuGlnLysTyrPheGluGlnHisLysThrAspLysValPro                               515520525                                                                      IleGlyThrValIleTyrThrThrMetGluProCysSerLeuArgLeu                               530535540                                                                      SerGlyAsnLysProCysValGluArgIleIleCysGlnGlnGlyAsn                               545550555560                                                                   IleThrAlaValPheValGlyValLeuGluProAspAsnPheValLys                               565570575                                                                      AsnAsnThrSerArgAlaLeuLeuGluGlnHisGlyIleAspTyrIle                               580585590                                                                      LeuValProGlyPheGlnGluGluCysThrGluAlaAlaLeuLysGly                               595600605                                                                      His                                                                            (2) INFORMATION FOR SEQ ID NO: 5:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 1082 base pairs                                                    (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: cDNA to mRNA                                               (iii) HYPOTHETICAL: NO                                                         (iii) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Ashbya gossypii                                                  (ix) FEATURE:                                                                  (A) NAME/KEY: 5'UTR                                                            (B) LOCATION: 1..314                                                           (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                              (B) LOCATION: 315..953                                                         (ix) FEATURE:                                                                  (A) NAME/KEY: 3'UTR                                                            (B) LOCATION: 954..1082                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:                                       CCCTTCTTGCACGGTCGTTTCTGAAACTCTACGATTATTGGAACAATGAGTAAGTCCTCA60                 AATGTACCACCTATCTGTAGTTTACTATCGGATTTACTGGCTAAGAGCTGACCTGTTAGG120                CAAGTGAAACATATCACATCGCCAGCAGGTTGGGCTACCAAGGATAGTTGATGACTTCCA180                TCACCTATAAAAGCGGCTTGAGTGCTTTTGCAATGATTCTGTTCACATGATGGACAAGAA240                ATACGTACAAAAATTTCAACGTTTTACAAGTTCCCAAGCTTAGTCAACTCATCACCAACG300                ACAAACCAAGCAACATGACAAGCCCATGCACTGATATCGGTACCGCTATA350                          MetThrSerProCysThrAspIleGlyThrAlaIle                                           1510                                                                           GAGCAGTTCAAGCAAAATAAGATGATCATCGTCATGGACCACATCTCG398                            GluGlnPheLysGlnAsnLysMetIleIleValMetAspHisIleSer                               152025                                                                         AGAGAAAACGAGGCCGATCTAATATGTGCAGCAGCGCACATGACTGCC446                            ArgGluAsnGluAlaAspLeuIleCysAlaAlaAlaHisMetThrAla                               303540                                                                         GAGCAAATGGCATTTATGATTCGGTATTCCTCGGGCTACGTTTGCGCT494                            GluGlnMetAlaPheMetIleArgTyrSerSerGlyTyrValCysAla                               45505560                                                                       CCAATGACCAATGCGATTGCCGATAAGCTAGACCTACCGCTCATGAAC542                            ProMetThrAsnAlaIleAlaAspLysLeuAspLeuProLeuMetAsn                               657075                                                                         ACATTGAAATGCAAGGCTTTCTCCGATGACAGACACAGCACTGCGTAT590                            ThrLeuLysCysLysAlaPheSerAspAspArgHisSerThrAlaTyr                               808590                                                                         ACAATCACCTGTGACTATGCGCACGGGACGACGACAGGTATCTCCGCA638                            ThrIleThrCysAspTyrAlaHisGlyThrThrThrGlyIleSerAla                               95100105                                                                       CGTGACCGGGCGTTGACCGTGAATCAGTTGGCGAACCCGGAGTCCAAG686                            ArgAspArgAlaLeuThrValAsnGlnLeuAlaAsnProGluSerLys                               110115120                                                                      GCTACCGACTTCACGAAGCCAGGCCACATTGTGCCATTGCGTGCCCGT734                            AlaThrAspPheThrLysProGlyHisIleValProLeuArgAlaArg                               125130135140                                                                   GACGGCGGCGTGCTCGAGCGTGACGGGCACACCGAAGCGGCGCTCGAC782                            AspGlyGlyValLeuGluArgAspGlyHisThrGluAlaAlaLeuAsp                               145150155                                                                      TTGTGCAGACTAGCGGGTGTGCCAGAGGTCGCTGCTATTTGTGAATTA830                            LeuCysArgLeuAlaGlyValProGluValAlaAlaIleCysGluLeu                               160165170                                                                      GTAAGCGAAAGGGACGTCGGGCTGATGATGACTTTGGATGAGTGTATA878                            ValSerGluArgAspValGlyLeuMetMetThrLeuAspGluCysIle                               175180185                                                                      GAATTCAGCAAGAAGCACGGTCTTGCCCTCATCACCGTGCATGACCTG926                            GluPheSerLysLysHisGlyLeuAlaLeuIleThrValHisAspLeu                               190195200                                                                      AAGGCTGCAGTTGCCGCCAAGCAGTAGACGGCAACGAGTTCTTTAAGTCGGTGT980                      LysAlaAlaValAlaAlaLysGln                                                       205210                                                                         TCATTTATGTAATATACCATTTCATCGAAAAAGTCAAATGGTATGAACTAGATTTATCAA1040               TAGTATCTAAGAGTTATGGTATTCGCAAAAGCTTATCGATAC1082                                 (2) INFORMATION FOR SEQ ID NO: 6:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 212 amino acids                                                    (B) TYPE: amino acid                                                           (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: protein                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:                                       MetThrSerProCysThrAspIleGlyThrAlaIleGluGlnPheLys                               151015                                                                         GlnAsnLysMetIleIleValMetAspHisIleSerArgGluAsnGlu                               202530                                                                         AlaAspLeuIleCysAlaAlaAlaHisMetThrAlaGluGlnMetAla                               354045                                                                         PheMetIleArgTyrSerSerGlyTyrValCysAlaProMetThrAsn                               505560                                                                         AlaIleAlaAspLysLeuAspLeuProLeuMetAsnThrLeuLysCys                               65707580                                                                       LysAlaPheSerAspAspArgHisSerThrAlaTyrThrIleThrCys                               859095                                                                         AspTyrAlaHisGlyThrThrThrGlyIleSerAlaArgAspArgAla                               100105110                                                                      LeuThrValAsnGlnLeuAlaAsnProGluSerLysAlaThrAspPhe                               115120125                                                                      ThrLysProGlyHisIleValProLeuArgAlaArgAspGlyGlyVal                               130135140                                                                      LeuGluArgAspGlyHisThrGluAlaAlaLeuAspLeuCysArgLeu                               145150155160                                                                   AlaGlyValProGluValAlaAlaIleCysGluLeuValSerGluArg                               165170175                                                                      AspValGlyLeuMetMetThrLeuAspGluCysIleGluPheSerLys                               180185190                                                                      LysHisGlyLeuAlaLeuIleThrValHisAspLeuLysAlaAlaVal                               195200205                                                                      AlaAlaLysGln                                                                   210                                                                            (2) INFORMATION FOR SEQ ID NO: 7:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 996 base pairs                                                     (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: cDNA to mRNA                                               (iii) HYPOTHETICAL: NO                                                         (iii) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Ashbya gossypii                                                  (ix) FEATURE:                                                                  (A) NAME/KEY: 5'UTR                                                            (B) LOCATION: 1..270                                                           (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                              (B) LOCATION: 271..789                                                         (ix) FEATURE:                                                                  (A) NAME/KEY: 3'UTR                                                            (B) LOCATION: 790..996                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7:                                       TGGTATAATGATACAGGAAGTGAAAATCCGAAAGGTTCAGACGATGAAAAGAGTTTGAGA60                 CGCATCAATGATCAGCTTTGAGCTATATGTAAGTCTATTAATTGATTACTAATAGCAATT120                TATGGTATCCTCTGTTCTGCATATCGACGGTTCTCACGTGATGATCAGCTTGAGGCTTCG180                CGGATAAAGTTCCATCGATTACTATAAAACCATCACATTAAACGTTCACTATAGGCATAC240                ACACAGACTAAGTTCAAGTTAGCAGTGACAATGATTAAGGGATTAGGCGAAGTT294                      MetIleLysGlyLeuGlyGluVal                                                       15                                                                             GATCAAACCTACGATGCGAGCTCTGTCGAGGTTGGCATTGTCCACGCG342                            AspGlnThrTyrAspAlaSerSerValGluValGlyIleValHisAla                               101520                                                                         AGATGGAACAAGACTGTCATTGACGCTCTCGACCAAGGTGCAATTGAG390                            ArgTrpAsnLysThrValIleAspAlaLeuAspGlnGlyAlaIleGlu                               25303540                                                                       AAACTGCTTGCTATGGGAGTGAAGGAGAAGAATATCACTGTAAGCACC438                            LysLeuLeuAlaMetGlyValLysGluLysAsnIleThrValSerThr                               455055                                                                         GTTCCAGGTGCGTTTGAACTACCATTTGGCACTCAGCGGTTTGCCGAG486                            ValProGlyAlaPheGluLeuProPheGlyThrGlnArgPheAlaGlu                               606570                                                                         CTGACCAAGGCAAGTGGCAAGCATTTGGACGTGGTCATCCCAATTGGA534                            LeuThrLysAlaSerGlyLysHisLeuAspValValIleProIleGly                               758085                                                                         GTCCTGATCAAAGGCGACTCAATGCACTTTGAATATATATCAGACTCT582                            ValLeuIleLysGlyAspSerMetHisPheGluTyrIleSerAspSer                               9095100                                                                        GTGACTCATGCCTTAATGAACCTACAGAAGAAGATTCGTCTTCCTGTC630                            ValThrHisAlaLeuMetAsnLeuGlnLysLysIleArgLeuProVal                               105110115120                                                                   ATTTTTGGTTTGCTAACGTGTCTAACAGAGGAACAAGCGTTGACACGT678                            IlePheGlyLeuLeuThrCysLeuThrGluGluGlnAlaLeuThrArg                               125130135                                                                      GCAGGCCTCGGTGAATCTGAAGGCAAGCACAACCACGGTGAAGACTGG726                            AlaGlyLeuGlyGluSerGluGlyLysHisAsnHisGlyGluAspTrp                               140145150                                                                      GGTGCTGCTGCCGTGGAGATGGCTGTAAAGTTTGGCCCACGCGCCGAA774                            GlyAlaAlaAlaValGluMetAlaValLysPheGlyProArgAlaGlu                               155160165                                                                      CAAATGAAGAAGTGAATATTAAAAAATCACTACTTAAAATTAACGTTTTTAT826                        GlnMetLysLys                                                                   170                                                                            TATGTCTATATCAAATTCTTACGTGATAACTTTTGATTTCGCTTCCTGGATTGGCGCAAG886                GCCTCCCTGTGTCGCAGTTTTTGTTCACGGGTCCACACAGCTCTGTTTTCCCAGAACATA946                TCCTCCCAGCCGGCGAACCGGTTAGACGCTTCTGCTGGCGTTCTTATTTT996                          (2) INFORMATION FOR SEQ ID NO: 8:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 172 amino acids                                                    (B) TYPE: amino acid                                                           (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: protein                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8:                                       MetIleLysGlyLeuGlyGluValAspGlnThrTyrAspAlaSerSer                               151015                                                                         ValGluValGlyIleValHisAlaArgTrpAsnLysThrValIleAsp                               202530                                                                         AlaLeuAspGlnGlyAlaIleGluLysLeuLeuAlaMetGlyValLys                               354045                                                                         GluLysAsnIleThrValSerThrValProGlyAlaPheGluLeuPro                               505560                                                                         PheGlyThrGlnArgPheAlaGluLeuThrLysAlaSerGlyLysHis                               65707580                                                                       LeuAspValValIleProIleGlyValLeuIleLysGlyAspSerMet                               859095                                                                         HisPheGluTyrIleSerAspSerValThrHisAlaLeuMetAsnLeu                               100105110                                                                      GlnLysLysIleArgLeuProValIlePheGlyLeuLeuThrCysLeu                               115120125                                                                      ThrGluGluGlnAlaLeuThrArgAlaGlyLeuGlyGluSerGluGly                               130135140                                                                      LysHisAsnHisGlyGluAspTrpGlyAlaAlaAlaValGluMetAla                               145150155160                                                                   ValLysPheGlyProArgAlaGluGlnMetLysLys                                           165170                                                                         (2) INFORMATION FOR SEQ ID NO: 9:                                              (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 1511 base pairs                                                    (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: cDNA to mRNA                                               (iii) HYPOTHETICAL: NO                                                         (iii) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Ashbya gossypii                                                  (ix) FEATURE:                                                                  (A) NAME/KEY: 5'UTR                                                            (B) LOCATION: 1..524                                                           (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                              (B) LOCATION: 525..1232                                                        (ix) FEATURE:                                                                  (A) NAME/KEY: 3'UTR                                                            (B) LOCATION: 1233..1511                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:                                       TGTATTCAACCTGGAGGATAACGAAATTTCCATGGCGCGGGCGATACCAACCCACAGGAG60                 CCAGATATAAGACCAATCCCGGCGGGTGTGCCAGCCGCCATCAGAGACAGCGGGCCAGCA120                AGGCATGTGAAGTCAAAAGGCGCCAGCTCCTTATCCGCTCCCGCACAAGCAGGACCGGCA180                TATCCCGATGAGCGCGCCAGCACCCAGACGCTACACCACCATTCGAAGTAGACTTTAAAA240                GAGCGCTTTCCAGCTTCTCAGGCAGTTAGCTCTACGACAAAGGAACCAAGTGATTTTCCC300                GATAGACGCGACTTGCTCAACGATGTTTCTGTGACCAGCGCAAGGAGAGATAGTCCTAAA360                GTATAATCAGATAGTTAGTCGTATCTTCTAGTTTTATTAGTCAGCTACATGGCGAACCGC420                CATTTCCTTATGCATGTCTTACGAGTTTAAAAAGCTCGCGGTAGCAGAAAAGAAGATGCA480                TAGATGGCATACCGAAGCCTATATCGCCCATAGAAGTTGATAGGATGTTTACCGGT536                    MetPheThrGly                                                                   ATAGTGGAACACATTGGCACTGTTGCTGAGTACTTGGAGAACGATGCC584                            IleValGluHisIleGlyThrValAlaGluTyrLeuGluAsnAspAla                               5101520                                                                        AGCGAGGCAGGCGGCAACGGTGTGTCAGTCCTTATCAAGGATGCGGCT632                            SerGluAlaGlyGlyAsnGlyValSerValLeuIleLysAspAlaAla                               253035                                                                         CCGATACTGGCGGATTGCCACATCGGTGACTCGATTGCATGCAATGGT680                            ProIleLeuAlaAspCysHisIleGlyAspSerIleAlaCysAsnGly                               404550                                                                         ATCTGCCTGACGGTGACGGAGTTCACGGCCGATAGCTTCAAGGTCGGG728                            IleCysLeuThrValThrGluPheThrAlaAspSerPheLysValGly                               556065                                                                         ATCGCACCAGAAACAGTTTATCGGACGGAAGTCAGCAGCTGGAAAGCT776                            IleAlaProGluThrValTyrArgThrGluValSerSerTrpLysAla                               707580                                                                         GGCTCCAAGATCAACCTAGAAAGGGCCATCTCGGACGACAGGCGCTAC824                            GlySerLysIleAsnLeuGluArgAlaIleSerAspAspArgArgTyr                               859095100                                                                      GGCGGGCACTACGTGCAGGGCCACGTCGACTCGGTGGCCTCTATTGTA872                            GlyGlyHisTyrValGlnGlyHisValAspSerValAlaSerIleVal                               105110115                                                                      TCCAGAGAGCACGACGGGAACTCTATCAACTTTAAGTTTAAACTGCGC920                            SerArgGluHisAspGlyAsnSerIleAsnPheLysPheLysLeuArg                               120125130                                                                      GATCAAGAGTACGAGAAGTACGTAGTAGAAAAGGGTTTTGTGGCGATC968                            AspGlnGluTyrGluLysTyrValValGluLysGlyPheValAlaIle                               135140145                                                                      GACGGTGTGTCGCTGACTGTAAGCAAGATGGATCCAGATGGCTGTTTC1016                           AspGlyValSerLeuThrValSerLysMetAspProAspGlyCysPhe                               150155160                                                                      TACATCTCGATGATTGCACACACGCAGACCGCTGTAGCCCTTCCACTG1064                           TyrIleSerMetIleAlaHisThrGlnThrAlaValAlaLeuProLeu                               165170175180                                                                   AAGCCGGACGGTGCCCTCGTGAACATAGAAACGGATGTTAACGGCAAG1112                           LysProAspGlyAlaLeuValAsnIleGluThrAspValAsnGlyLys                               185190195                                                                      CTAGTAGAGAAGCAGGTTGCACAGTACCTGAATGCGCAGCTGGAAGGT1160                           LeuValGluLysGlnValAlaGlnTyrLeuAsnAlaGlnLeuGluGly                               200205210                                                                      GAGAGCTCGCCATTGCAGCGCGTGCTCGAAAGGATTATTGAATCCAAG1208                           GluSerSerProLeuGlnArgValLeuGluArgIleIleGluSerLys                               215220225                                                                      CTTGCTAGCATCTCAAATAAGTGATTATATTATCTTGGGTGCTGTATATCT1259                        LeuAlaSerIleSerAsnLys                                                          230235                                                                         TATGTATGTCTTACGACTGTGAATCAGAGGGGTGGCAGCTGGAACACCAGCGACACACCT1319               TCGTCTCCCGCGGTGATCAGCCTTCTGTTTTCCTCAAGTAGTACAAAGTCTAGGACACCC1379               TGTTGTGGCCAACGCAAACATGGAGCTGCTGCCCGTTACGCACGTCGAACTCGTAGACCT1439               TGCCGTCAATGCACGAGGCGAACAGGTGGAAACCGGTGGTCTTGTCAAACCGCCAGCTTC1499               GTGACCGAGTCC1511                                                               (2) INFORMATION FOR SEQ ID NO: 10:                                             (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 235 amino acids                                                    (B) TYPE: amino acid                                                           (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: protein                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10:                                      MetPheThrGlyIleValGluHisIleGlyThrValAlaGluTyrLeu                               151015                                                                         GluAsnAspAlaSerGluAlaGlyGlyAsnGlyValSerValLeuIle                               202530                                                                         LysAspAlaAlaProIleLeuAlaAspCysHisIleGlyAspSerIle                               354045                                                                         AlaCysAsnGlyIleCysLeuThrValThrGluPheThrAlaAspSer                               505560                                                                         PheLysValGlyIleAlaProGluThrValTyrArgThrGluValSer                               65707580                                                                       SerTrpLysAlaGlySerLysIleAsnLeuGluArgAlaIleSerAsp                               859095                                                                         AspArgArgTyrGlyGlyHisTyrValGlnGlyHisValAspSerVal                               100105110                                                                      AlaSerIleValSerArgGluHisAspGlyAsnSerIleAsnPheLys                               115120125                                                                      PheLysLeuArgAspGlnGluTyrGluLysTyrValValGluLysGly                               130135140                                                                      PheValAlaIleAspGlyValSerLeuThrValSerLysMetAspPro                               145150155160                                                                   AspGlyCysPheTyrIleSerMetIleAlaHisThrGlnThrAlaVal                               165170175                                                                      AlaLeuProLeuLysProAspGlyAlaLeuValAsnIleGluThrAsp                               180185190                                                                      ValAsnGlyLysLeuValGluLysGlnValAlaGlnTyrLeuAsnAla                               195200205                                                                      GlnLeuGluGlyGluSerSerProLeuGlnArgValLeuGluArgIle                               210215220                                                                      IleGluSerLysLeuAlaSerIleSerAsnLys                                              225230235                                                                      (2) INFORMATION FOR SEQ ID NO: 11:                                             (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 1596 base pairs                                                    (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: double                                                       (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: cDNA to mRNA                                               (iii) HYPOTHETICAL: NO                                                         (iii) ANTI-SENSE: NO                                                           (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Ashbya gossypii                                                  (ix) FEATURE:                                                                  (A) NAME/KEY: 5'UTR                                                            (B) LOCATION: 1..352                                                           (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                              (B) LOCATION: 353..1093                                                        (ix) FEATURE:                                                                  (A) NAME/KEY: 3'UTR                                                            (B) LOCATION: 1094..1596                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:                                      AGAAGAAGCGCAGGCGCCAGTCCGAGCTGGAGGAGAACGAGGCGGCGCGGTTGACGAACA60                 GCGCGCTGCCCATGGACGATGCGGGTATACAGACGGCGGGTATACAGACGGCGGGTGGTG120                CCGAGAGAGGCACCAGGCCGGCTTCCTCCAGCGATGCAAGGAAGAGAAGGGGACCAGAGG180                CGAAGTTCAAGCCATCTAAGGTACAGAAGCCCCAATTGAAGCGAACTGCATCGTCCCGGG240                CGGATGAGAACGAGTTCTCGATATTATAGAGGCCCCCGTTTCGAGTGATTGGCGTCAAAA300                ACGGCTATCTGCCTTCGTCCGCCCCCACCACCCTCGGGAACACTGGCAAACCATG355                     Met                                                                            1                                                                              GCGCTAATACCACTTTCTCAAGATCTGGCTGATATACTAGCACCGTAC403                            AlaLeuIleProLeuSerGlnAspLeuAlaAspIleLeuAlaProTyr                               51015                                                                          TTACCGACACCACCGGACTCATCCGCACGCCTGCCGTTTGTCACGCTG451                            LeuProThrProProAspSerSerAlaArgLeuProPheValThrLeu                               202530                                                                         ACGTATGCGCAGTCCCTAGATGCTCGTATCGCGAAGCAAAAGGGTGAA499                            ThrTyrAlaGlnSerLeuAspAlaArgIleAlaLysGlnLysGlyGlu                               354045                                                                         AGGACGGTTATTTCGCATGAGGAGACCAAGACAATGACGCATTATCTA547                            ArgThrValIleSerHisGluGluThrLysThrMetThrHisTyrLeu                               50556065                                                                       CGCTACCATCATAGCGGCATCCTGATTGGCTCGGGCACAGCCCTTGCG595                            ArgTyrHisHisSerGlyIleLeuIleGlySerGlyThrAlaLeuAla                               707580                                                                         GACGACCCGGATCTCAATTGCCGGTGGACACCTGCAGCGGACGGGGCG643                            AspAspProAspLeuAsnCysArgTrpThrProAlaAlaAspGlyAla                               859095                                                                         GATTGCACCGAACAGTCTTCACCACGACCCATTATCTTGGATGTTCGG691                            AspCysThrGluGlnSerSerProArgProIleIleLeuAspValArg                               100105110                                                                      GGCAGATGGAGATACCGCGGGTCCAAAATAGAGTATCTGCATAACCTT739                            GlyArgTrpArgTyrArgGlySerLysIleGluTyrLeuHisAsnLeu                               115120125                                                                      GGCAAGGGGAAGGCGCCCATAGTGGTCACGGGGGGTGAGCCGGAGGTC787                            GlyLysGlyLysAlaProIleValValThrGlyGlyGluProGluVal                               130135140145                                                                   CGCGAACTAGGCGTCAGTTACCTGCAGCTGGGTGTCGACGAGGGTGGC835                            ArgGluLeuGlyValSerTyrLeuGlnLeuGlyValAspGluGlyGly                               150155160                                                                      CGCTTGAATTGGGGCGAGTTGTTTGAGCGACTCTATTCTGAGCACCAC883                            ArgLeuAsnTrpGlyGluLeuPheGluArgLeuTyrSerGluHisHis                               165170175                                                                      CTGGAAAGTGTCATGGTCGAAGGCGGCGCGGAGGTGCTCAACCAGCTG931                            LeuGluSerValMetValGluGlyGlyAlaGluValLeuAsnGlnLeu                               180185190                                                                      CTGCTGCGCCCAGATATTGTGGACAGTCTGGTGATCACGATAGGATCC979                            LeuLeuArgProAspIleValAspSerLeuValIleThrIleGlySer                               195200205                                                                      AAGTTCCTGGGCTCACTAGGTGTTGCGGTCTCACCAGCTGAGGAGGTG1027                           LysPheLeuGlySerLeuGlyValAlaValSerProAlaGluGluVal                               210215220225                                                                   AACCTAGAGCATGTGAACTGGTGGCACGGAACAAGTGACAGTGTTTTG1075                           AsnLeuGluHisValAsnTrpTrpHisGlyThrSerAspSerValLeu                               230235240                                                                      TGCGGCCGGCTCGCATAGCGGTTATGACTGGTCTACTAGTTAAAACTATTTACTC1130                    CysGlyArgLeuAla                                                                245                                                                            CTATACATATTGCGTCACATAGCGTTTATCCCCCTCGCCAACCGCCTCGTGCCGTTGGAA1190               ACACGGCGGCCGGGGGACCTCAAGCGCTCCGCATCGACTAGTTTAATTTACAAACAGATT1250               CTGTAACTTGCGTAACGGCCAGAGGTCTCTGACTTTCTGATAATCTTCACCACCTCACCT1310               CGCTTCAACCCCAGGTATAATGCAACTTGGATCCATCCTCTGGATTCTAGGTAACTGAGA1370               TTCCTTTAACCTGTATCTCTTCAACAACTCCTTCTTTTCTTCGTCGCTGAGTTTGATATG1430               TTTTGGCACAAGCTCATGGTGCGTGATATTTACCACCAAAGCTGTTTCGTTGAAAGTCTC1490               AATTGTAGCAGGAGCGACGGAGGGAAGCAGTTTCAACGCGCTGGGCGTTATGCCGTTCTG1550               ATATATGAAAATACCCGTCTGGAAGTTCTTCTCGCCAATGTGGATC1596                             (2) INFORMATION FOR SEQ ID NO: 12:                                             (i) SEQUENCE CHARACTERISTICS:                                                  (A) LENGTH: 246 amino acids                                                    (B) TYPE: amino acid                                                           (D) TOPOLOGY: linear                                                           (ii) MOLECULE TYPE: protein                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12:                                      MetAlaLeuIleProLeuSerGlnAspLeuAlaAspIleLeuAlaPro                               151015                                                                         TyrLeuProThrProProAspSerSerAlaArgLeuProPheValThr                               202530                                                                         LeuThrTyrAlaGlnSerLeuAspAlaArgIleAlaLysGlnLysGly                               354045                                                                         GluArgThrValIleSerHisGluGluThrLysThrMetThrHisTyr                               505560                                                                         LeuArgTyrHisHisSerGlyIleLeuIleGlySerGlyThrAlaLeu                               65707580                                                                       AlaAspAspProAspLeuAsnCysArgTrpThrProAlaAlaAspGly                               859095                                                                         AlaAspCysThrGluGlnSerSerProArgProIleIleLeuAspVal                               100105110                                                                      ArgGlyArgTrpArgTyrArgGlySerLysIleGluTyrLeuHisAsn                               115120125                                                                      LeuGlyLysGlyLysAlaProIleValValThrGlyGlyGluProGlu                               130135140                                                                      ValArgGluLeuGlyValSerTyrLeuGlnLeuGlyValAspGluGly                               145150155160                                                                   GlyArgLeuAsnTrpGlyGluLeuPheGluArgLeuTyrSerGluHis                               165170175                                                                      HisLeuGluSerValMetValGluGlyGlyAlaGluValLeuAsnGln                               180185190                                                                      LeuLeuLeuArgProAspIleValAspSerLeuValIleThrIleGly                               195200205                                                                      SerLysPheLeuGlySerLeuGlyValAlaValSerProAlaGluGlu                               210215220                                                                      ValAsnLeuGluHisValAsnTrpTrpHisGlyThrSerAspSerVal                               225230235240                                                                   LeuCysGlyArgLeuAla                                                             245                                                                            __________________________________________________________________________ 

We claim:
 1. An isolated DNA sequence which codes for a polypeptide with the amino acid sequence (GTP cyclohydrolase II) depicted in SEQ ID NO: 2, or for an analog or derivative of the polypeptide depicted in SEQ ID NO: 2 having essentially the same enzymatic activity as the polypeptide depicted in SEQ ID NO: 2, with the proviso that the DNA sequence of the analog or derivative of the polypeptide depicted in SEQ ID NO: 2 hybridizes with the polynucleotide depicted in SEQ ID NO: 1 at 42° C. in 5×SSC in 50% formamide solution.
 2. An isolated DNA sequence which codes for a polypeptide with the amino acid sequence (DRAP deaminase) depicted in SEQ ID NO: 4, or for an analog or derivative of the polypeptide depicted in SEQ ID NO: 4 having essentially the same enzymatic activity as the polypeptide depicted in SEQ ID NO: 4, with the proviso that the DNA sequence of the analog or derivative of the polypeptide depicted in SEQ ID NO: 4 hybridizes with the polynucleotide depicted in SEQ ID NO: 3 at 42° C. in 5×SSC in 50% formamide solution.
 3. An isolated DNA sequence which codes for a polypeptide with the amino acid sequence (DBP synthase) depicted in SEQ ID NO: 6, or for an analog or derivative of the polypeptide depicted in SEQ ID NO: 6 having essentially the same enzymatic activity as the polypeptide depicted in SEQ ID NO: 6, with the proviso that the DNA sequence of the analog or derivative of the polypeptide depicted in SEQ ID NO: 6 hybridizes with the polynucleotide depicted in SEQ ID NO: 5 at 42° C. in 5×SSC in 50% formamide solution.
 4. An isolated DNA sequence which codes for a polypeptide with the amino acid sequence (DMRL synthase) depicted in SEQ ID NO: 8, or for an analog or derivative of the polypeptide depicted in SEQ ID NO: 8 having essentially the same enzymatic activity as the polypeptide depicted in SEQ ID NO: 8, with the proviso that the DNA sequence of the analog or derivative of the polypeptide depicted in SEQ ID NO: 8 hybridizes with the polynucleotide depicted in SEQ ID NO: 7 at 42° C. in 5×SSC in 50% formamide solution.
 5. An isolated DNA sequence which codes for a polypeptide with the amino acid sequence (riboflavin synthase) depicted in SEQ ID NO: 10, or for an analog or derivative of the polypeptide depicted in SEQ ID NO: 10 having essentially the same enzymatic activity as the polypeptide depicted in SEQ ID NO: 10, with the proviso that the DNA sequence of the analog or derivative of the polypeptide depicted in SEQ ID NO:10 hybridizes with the polynucleotide depicted in SEQ ID NO: 9 at 42° C. in 5×SSC in 50% formamide solution.
 6. An isolated DNA sequence which codes for a polypeptide with the amino acid sequence (HTP reductase) depicted in SEO ID NO: 12, or for an analog or derivative of the polypeptide depicted in SEQ ID NO: 12 having essentially the same enzymatic activity as the polypeptide depicted in SEQ ID NO: 12, with the proviso that the DNA sequence of the analog or derivative of the polypeptide depicted in SEQ ID NO: 12 hybridizes with the polynucleotide depicted in SEQ ID NO: 11 at 42° C. in 5×SSC in 50% formamide solution.
 7. An expression vector containing one or more DNA sequences as defined in any one of the preceeding claims.
 8. A host organism which has been transformed with an expression vector as defined in claim
 7. 9. A recombinant method of producing riboflavin, comprising the steps of1) constructing the expression vector defined in claim 7; 2) transforming a compatible host with said recombinant vector such that the DNA sequence coding for the polypeptide can be expressed by the host; 3) culturing the transformed host in a suitable growth medium to produce said riboflavin; and 4) recovering said riboflavin from the growth medium. 